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- BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Study Title Hydrolysis Reactionsof 2-(N-MethylperfluoroctanesuIfonamido)-EthylAlcohol (N-MeFOSE Alcohol) Data Requirement: Based on OPPTS:835.2110 Author Thomas L. Hatfield, Ph.D. Study Completion Date March 30,2001 Performing Laboratory 3M Environmental Laboratory Building2-3E-09,935 Bush Avenue St. Paul, MN 55106 Project Identification 3M Laboratory Report No: W1880 Total Number of Pages 71 Page 1 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 This page has been reserved for specific country requirements. Page 2 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Statement of Non-Compliance Study Title: Hydrolysis Reactions of 2-(N-Methylperfiuoroctanesu1fonamido)-Ethyl Alcohol (N-MeFOSE Alcohol) Study Identification Number: W1880 This study does not fully comply with the requirements of the US EPA Good Laboratory Practices (GLP) Standards at 40 CFR Part 792 (TSCA). However, many GLP standards were used in the development of the analytical method (Appendix A), and the quality assurance proceduresfollowed in this study were based on the practicesdescribed in the GLP documentation. This is a revised report in that the statistics on the study data and the discussion were changed from the initial study report. Changes to these interpretive sections were made to better represent the experimental results of the study. fl-/&u Study Director *//& r- Sponsor Representative Date Page 3 of 71 ~~ BACK TO MAIN 3M Environmental Laboratory Report No.W1880 ~~~~ Quality Assurance Statement Study Title: Hydrolysis Reactions of 2-(N-Methylperfluoroctanesulfonamido)-Ethyl Alcohol (N-MeFOSE Alcohol) Study Identification Number: W1880 The following table provides details of the audits performed by the 3M Environmental Laboratory Quality Assurance Unit (QAU). Inspection Dates Phase Date Reported to Management Study Director I I I I 10/4-6/2000 II I I I I 3/19, 20/2001 Data and Draft Report Draft Report I 10/06/00 3/21/01 10/06/00 3/121/01 / Date 2 1 2001 Page 4 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Table of Contents Statement of Non-Compliance ............................................................................................ 3 Quality Assurance Statement .............................................................................................. 4 List of Tables ........................................................................................................................ 6 List of Figures....................................................................................................................... 6 Study Personnel and Contributors....................................................................................... 6 Location of Archives ............................................................................................................. 7 Summary.............................................................................................................................. 8 Introduction .......................................................................................................................... 9 Summary of Kinetics Model............................................................................................... 10 Materials and Methods....................................................................................................... 11 Chemical Characterizations.......................................................................................... 11 Sample Preparation ...................................................................................................... 11 Sample Analysis............................................................................................................ 12 Deviations ...................................................................................................................... 12 Results and Discussion ..................................................................................................... 14 Data Quality Objectives (DQO's) .................................................................................. 14 Anomalous Analytical Results....................................................................................... 14 Statistical Methods and Calculations ............................................................................ 15 Data Summary and Discussion..................................................................................... 15 Conclusions........................................................................................................................ 19 References......................................................................................................................... 20 Signatures .......................................................................................................................... 21 Appendix A: Analytical Method .......................................................................................... 22 Appendix B: Kinetics Model............................................................................................... 39 Appendix C: Selected Analytical and Kinetics Results ..................................................... 49 Appendix D::Selected Chromatograms............................................................................. 60 Page 5 of 71 BACK TO MAIN 3M EnvironmentalLaboratory Report No. W1880 List of Tables Table 1. Summary of Results Based on N-MeFOSEAlcohol Concentrations..................8 Table 2. Summary of Results Based on PFOS Limit of Quantification ............................. 8 Table 3. Characterizationsof Test and Reference Substances ...................................... 11 Table 4. Observed (50"C) Degradation Rates of N-MeFOSE Alcohol in Aqueous Buffered Solutions and at Various pH Levels..................................................... 15 Table 5. Degradation Rate and Half Life of N-MeFOSE Alcohol in Aqueous Buffered Solutions Using Data Pooled Over pH Levels.................................................... 16 Table 6. Degradation Rate and Half Life of N-MeFOSEAlcohol in Aqueous Buffered Solutions Based on PFOS Limit of Quantification.............................................. 18 List of Figures Figure 1. Structures of N-MeFOSE Alcohol and the Potassium Salt of PFOS................9 Figure 2. Observed N-MeFOSE Alcohol Degradation for Various pH Levels .................16 Figure 3. Pooled N-MeFOSE Alcohol Data and Slope Regression. ................................ 17 Study Personnel and Contributors Study Director Thomas L. Hatfield, Ph.D. 3M Environmental Laboratory Building 2-3E:-09 935 Bush Avenue St. Paul, MN 55106 (651) 778-7863 Sponsor 3M Corporation 3M Environmental Laboratory and Professional Services Contributing Personnel Kuruppu Dharmasiri, Ph.D Mark T. McCann Anthony E. Scales Joseph J. S. Tokos, Ph.D (Pace Analytical Services, Inc., 1700 Elm St., Minneapolis, MN 55144) Gregory Maisel Jill Maloney (Braun Intertec Corporation, 6875 Washington Ave. South, Minneapolis, MN 55439) Page 6 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Location of Archives The 3M Environmental Laboratory will retain the original data documents and digital copies of the original data related to this work for at least 10 years following the effective date of any related final ruling. Information may be obtained through written inquiry addressed as follows: 3M Environmental Laboratory Building 2-3E-09 935 Bush Avenue St. Paul, MN 55106 Page 7 of 71 BACK TO MAIN 3M EnvironmentalLaboratory Report No. W1880 Summary We report here the results of our study of the hydrolysis of 2-(N-methylperfluorooctanesulfonamid0)-ethylalcohol (hereafter, N-MeFOSEAlcohol). Our methods are described below and in Appendix A to this work; our results are based on the observed concentrations of N-MeFOSE alcohol and its potential hydrolysis product perfluorooctane sulfonate (PFOS) in buffered aqueous solutions as a function of time. The chosen analytical technique was high performance liquid chromatography with mass spectrometry detection (HPLUMS). Tables 1 and 2 summarize the results of the study. During this study, we prepared and examined samples at six different pH levels from 1.5 to 11.Oover a period of 49 days, and our results indicate no dependence of the degradation rate of N-MeFOSE alcohol on the sample pH level. Our results based on the N-MeFOSE alcohol concentrations,pooled over the observed pH levels, are presented in Table 1. Table 1. Summary of Results Based on N-MeFOSE Alcohol I Concentrations I Observed Rate Calculated I I I I I I (years) at 25" C (years) I I I I I 0.0030 0.00030 I 6.3 I 3.8 to 19.4 We also monitored the concentration of one of the potential hydrolysis products (perfluorooctane sulfonate, PFOS), but never observed this compound at levels above its limit of quantification (LOQ, equal to 12.8 ng/mL). Assuming PFOS to be the only hydrolysis product of N-MeFOSE Alcohol, this LOQ (and other experimental data) provide a second estimate of the N-MeFOSE alcohol half-life, presented in Table 2. I Table 2. Summary of Results Based on PFOS Limit of Quantification I Maximum Maximum Possible Rate Calculated Rate Calculated Half Life Constant at 50" C Constant at 25" C at 25" C (day') 7.0 x104 (day1) 7.0 XO-'I (years) 2 27 According to the data available from this study, the half-life estimate of Table 2 represents the minimum possible half-life of the compound N-MeFOSE alcohol under the assumption that it hydrolyzes to form only the compound PFOS. Page 8 of 71 BACK TO MAIN 3M EnvironmentalLaboratory Report No. W1880 I ntroduction Three primary chemical routes of environmental degradation are hydrolysis, photolysis, and biodegradation. Studies of these routes provide information on the environmental persistence of both the "parent" compounds and their reaction products, and are ideally carried out over the range of chemical conditions pertinent to both environmental and metabolic processes. The hydrolysis of N-MeFOSE alcohol (or, more generally, its degradation in the presence of %O) is addressed in this report. Structures of the "parent" compound N- MeFOSE alcohol and the potassium salt of possible hydrolysis product perfluorooctane sulfonate (PFOS) are illustrated in Figure 1. Figure 1. Structuresof N-MeFOSE Alcohol and the PotassiumSalt of PFOS N-MeFOSE Alcohol OH Potassium Salt of PFOS FllrTTlll IIS-0- K' FFFFFFFFO Page 9 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Summary of Kinetics Model A full mathematical description of the kinetics model employed in this study is presented in Appendix B. The study data allow two independent estimates of the hydrolytic half-life of N-MeFOSE Alcohol. The first estimate (see Table 1) is based on the observed degradation of the "parent" compound PJ-MeFOSE alcohol in dilute, appropriately buffered aqueous solutions. Equation 1 describes the estimated half-life(f'yi)i in terms of the estimated total parent hydrolysis rate i p(see Appendix B, Equation BIO): w= - 2) kP Eq. 1 We determined the quantity c pfrom the experimentaldata as described in Appendix B. At each pH level, we used the earliest study data meeting the data quality objectives to determine the relevant concentration ratios (see Equation B8). The measured concentrations of the potential hydrolysis product PFOS (also obtained during the experiments described here) provide a second estimate (see Table 2) of the parent half-life. During the course of this study, we did not detect PFOS above its limit of quantitation (LOQ), and related studies' show that PFOS is itself hydrolytically stable. vi)2 Assuming also that PFOS is the only hydrolytic product of the parent compound N- MeFOSE alcohol, these PFOS analyses provide the following estimate (T of the minimum N-MeFOSE alcohol half-life (see Equations 832 and B33): Eq. 2 where [Po]= A t= the initial N-MeFOSE alcohol molar concentration, the time interval over which the study was conducted (49 days), and = the molar limit of quantitation for the compound PFOS. All the samples used in this study were maintainedat a reactiontemperature of 50"C. The quoted results, valid for the reaction temperature of 25" C, were calculated from our experimental results according to methods described in Appendix B (Eq. B38 and B39). Page 10 of 71 BACK TO MAIN 3M EnvironmentalLaboratory Report No. W1880 ____ Materials and Methods Details of the characteristics of the test materials, sample preparationtechniques, and analytical methods are presented in Appendix A (ETS-8-178.0, "Preparation of 2-(NMethylperfluoroctanesulfonamido)Ethyl Alcohol (N-MeFOSE Alcohol) Hydrolysis Samples and Analysis by High PerformanceLiquid Chromatographywith Mass Spectrometry Detection.") A summary of these items is provided below, as well as a description the known deviations from the procedures of Appendix A. 3M prepared and analyzed the samples included in this study between March 13 and October 5, 1999. Chemical Characteizations Table 3 describes the sources and properties of the materials used in this work. These materials were used to prepare both the samples and the quantitative standards used to quantify them. For this reason, and because Equation 88 (see Appendix B) involves only ratios of parent concentrations,the resulting rate and half-lifeestimates are largely independent of the material purity levels. Table 3. Characterizationsof Test and Reference Substances Source Chemical Lot Numbee N-EtFOSE Alcohol` 3M Specialty Chemistry TN-A1284 Physical Description Molecular Weight (gm mole-`) Off-white powder 570.9 PFOS 3CMhemistry TN-A2130 SD108 White powder 498.9 1 THPFOS~ ICN Biornedicals TN-A1339 (Lot #59909, SD028) Light brown powder 428.0 N-MeFOSE Alcohol 3M ICp/PCP Division TN-A1282 SD 015 Amber waxy solid 556.9 a The "S" and "TNA" designations are based on reference numbers in two redundant databases maintained by 3M. b 3, 3, 4, 4, 5,5,6,6,7, 7, 8, 8, atridecafluorooctane sulfonic acid. 2-(Kethylpeifluoroctanesulfonamido)-ethylalcohol. Sample Preparation We prepared three 1.O-mL aqueous buffer samples (a sample, a duplicate, and a "matrix spike") at each of six pH levels (1.5, 3, 5, 7, 9 and 11) for analysis at eight time intervals (0,7,14,21, 28,35,42 and 49 days). Bufferedsolutions containingthe analyte NMeFOSE alcohol and THPFOS (3, 3,4,4, 5, 5, 6,6, 7, 7, 8,8, 8-tridecafluorooctane sulfonic acid), the latter serving as a surrogate for the compound PFOS, formed the basis of all these samples. The chosen buffer solutions are described fully in Appendix A We prepared all the samples simultaneously,and placed all but the "Day 0 samples in an orbital inc:ubator/shakermaintainedat 50" (k3") C. After at least three minutes of agitation, we diluted the "Day 0 samples 1O:l with methanol, added solutions of the internal standard 2-(N-ethylperfluoroctanesulfonamido)-ethylalcohol (N-EtFOSE Page 11 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 alcohol), spiked the samples (as required) with a solution of N-MeFOSE alcohol, and then refrigerated them. After the appropriate incubation times, subsets of the sample vials were removed from the incubator and then diluted, spiked, and stored as described immediately above. Except during the relatively short periods of time required to prepare them, the samples were shielded from light. In all the samples, the resulting THPFOS and N-EtFOSE alcohol concentrationswere 403 and 31 1 ng/ml, respectively. In the samples spiked with the anlatye N-MeFOSE alcohol, the resulting analyte levels were 221 ng/ml above those in the unspiked samples. Six calibration standards containing N-MeFOSEalcohol (78.9 to 789 ng/ml), N-EtFOSE alcohol (311 ng/ml), THPFOS (403 ng/ml) and PFOS (3.1 to 94 ng/ml) served as the quantitative basis of the study. All these standards were prepared at the appropriate pH levels using the chosen buffer solutions (see Appendix A.) Samplehlysii The equipment we used for the HPLC/MS analysis was a Hewlett Packard model 1100 equipped with a Dionex lonPac@NG-1 HPLC column (aqueous ammonium acetate/methanolsolvent gradient) and an ALS Model G1322A degassing module. An ALS Model G1315A column heater maintainedthe column temperature at 40 "C, a quaternary pump supplied a column flow rate of 0.3 mumin, and an ALS Model G1313A auto-sampler provided 5 pL sample injections. The detector was a Hewlett Packard MSD mass spectrometer,operated in negative-modeelectrospray ionization mode; anions of PFOS, THPFOS, and acetate adducts of N-MeFOSE alcohol and N-MeFOSE alcohol were detected at the charge-to mass ratio (m/z) values 499,427, 616, and 630 respectively. We processed the resulting data using the computer program HP ChernStation for LC (Rev.A.06.0). The calibration curves and analytical results are based on the measured area ratios for the analytelinternal standard pairs; N-EtFOSE alcohol served as the internal standard for N-MeFOSE alcohol, and THPFOS served as the internal standard for PFOS. Additional analytical details, includingthe gradient elution program, instrument and detector parameters, and performance specifications, are presented in Appendix A. Deviations The gradient parameters and column temperature were incorrectly entered on the instrument plarameters spreadsheet dated 12117/99. The correct parameters are listed in the data acquisition method file "F0SESIM.m." Through either human or mechanical error, samples for "Day 42,"pH 1.5 were not analyzed. The mass spectrometer detector gain was set to 2.0 for samples analyzed on 12/20/99. Earlier sets were analyzed with gain = 1 .O. The pH 3 data were reanalyzed using the correct quantitative ion at m/z = 616; an earlier analysis incorrectly employed the "monitor" ion value m/z = 617. Page 12 of 71 BACK TO MAIN 3M Environmental Laboratory Report No.W1880 The actual concentration of the THPFOS solution added as a surrogate (21,030 CLg/mL) was incorrectly entered as 20,130 pg/mL in some documents. This error has no effect on the accuracy of the analytical results, but has been noted on the reagent preparation logs. The analytical resultsfor one set of samples, intendedfor the pH 1.5, "Day 4 9 analysis, indicated much lower concentrations of N-MeFOSE than for any other similar sample prepared for this study. All other analytical results were quite consistent, so it is highly likely that these samples were not properly prepared. Incorrect amounts of either the surrogate or internal standard solutions were added to the following pH = 1.5 samples: MFA-002, 127, 128, and 129. The same is true of the following pH = 11 samples: MFAO16,017,018,034, 035,036, 124, 125, and 126. In many cases, these samples were rejected on the basis of the data quality objectives (see the section immediately below). The resulting analytical values for the surrogate and internal standards were also excluded from evaluations of the consistency of the related results (see the "Data Summary and Discussion" section below). Page 13 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Results and Discussion Data Quality objectives(DaYs) Below is a brief description of the data quality objectives applied in this study. A full description is presented in Appendix A. With the exceptions of the anomalous results noted below, all the DQO's were met. Appendix C presents the results for each sample set, organized by pH level. Calibrations. The minimum acceptable coefficient of determination (?) for linear fits to calibration data is 0.990. The acceptance criterion for individual calibration points is that their values fall within f 25% of the linear fit value; data outside this range are excluded and the calibration curve is recalculated. No more than two points may be rejected from a calibration data set. Data for the high or low calibration standards may be rejected, though this results in a smaller effective calibration range. Continuing Calibration Verification (CCV). Identical calibration samples are examined at the beginning and end of each sample run. Results of the second calibration run may not deviate by more than f 25% of the first run for any analyte. The average results of the calibration runs are used to calculate the analyte concentrations. Matrix Spikes. The acceptable percent spike recovery range is 75% to 125%. Analyte specificity is demonstrated by acceptable analyte spike recoveries. Sample Duplicates. Duplicate pairs with relative percent deviation (RSD) greater than 25% may be accepted at the analyst`s discretion, but must be noted. Solvent Blanks. Concentration results for solvent blanks may exceed neither 5% of the highest calibration standard nor 25% of the lowest calibration level. System Suitability. Suitability was demonstrated by either an abbreviated mass-tocharge (m/z) check-tuneor performance of a full auto-tune routine. AnomalousAnalytical Resutts Calibrations. Of the 144 calibration results obtained, 13 individual values failed to meet the stated DQO and were rejected. We rejected fewer than two values for any particular calibration run and compound. Spike Recoveries. Results for the following sample pairs failed to meet this data quality objective and are excluded from the following analyses: MFA-127-128 ("Day 4 9 , pH = 1.5;);MFA-004-005 ("Day 0,pH = 3.0); MFA-016-017 ("Day 0,pH = 11); MFA034-035 ("Day 7", pH = II); and MFA-124-125 ("Day 4 2 , pH = 11). Sample Duplicates. The following sample pairs failed to meet this data quality objective and are excluded from the following analyses: MFA-001-002 ("Day 0, pH = 1.5)and MFA040-041 ("Day 14, pH = 3.0). Solvent Blanks. PFOS results for two of the 12 solvent blanks (the initial blanks performed at pH levels 3.0 and 7.0) exceeded 25% of the calibration standards values below 15.7 ng/ml. As a result, the detection limit for PFOS quoted in this study is '15.7 ng/ml (see Appendix A). Page 14 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Statistical Methods and Calculations Using functions provided in Microsoft@E'xcel@s'oftware, we calculated means, standard deviations, and first-order rate constants (see Appendix B, Equation B8) for various subsets of the acquired data. Our linear regressions included the determination of constant terms, that is, we did not force the regression fits to pass through the origin. As described in Appendix B (Equations B38 and B39), rates measured at 50C were extrapolated to 25C by dividing by a factor of 10; this approximation is valid for reactions with Arrhenius heats of activation near 18 Kcal/mole.* Data Summary and Discussion The LOQ is defined as the concentration of the lowest (accepted) standard in the calibration set for which the known concentration exceeds 400% of the indicated solvent blank level (see Appendix A). During this study, the LOQ's for N-MeFOSE alcohol and PFOS were 78.9 ng/mL and 15.7 ng/mL, respectively. Excluding those samples noted above in the "Deviation" section, our results for the surrogate compound (THPFOS) and the internal standard (N-EtFOSE alcohol) were quite consistent throughout the study. The percent relative standard deviations of the measured THPFOS values, calculated for each pH level, ranged from 2.1 % to 10.2%. The corresponding values for N-EtFOSE alcohol ranged from 4.7% to 11.3%. Figure 2 illustrates the observed N-MeFOSE alcohol concentrations at the six pH levels (at 50C),and Table 4 presentsthe results of the slope determinations (seeEquation 88) for the same data. ~ ~ Table 4. Observed (50" C) Degradation Rates of N-MeFOSEAlcohol in Aqueous Buffered Solutions and at Various pH Levels. ~ PH ~ Observed Rate (day') ~ ~~ ~ Percent (20) Rate Uncertainty 1.5 4.26 x IO-' 67 3.0 4.42x I O 4 582 5.0 5.68 1 0 . ~ 25 7.0 4.83x 10" 24 9.0 3.97 x 34 11 2.98 x 10" 25 These degradation rates, with the exception of the pH = 3.0 data, are generally well determined, with percent relative 20 (95% confidence) uncertainties in the range 24% to 67%. The data do not indicate any clear dependence of the degradation rate on the sample pH. Page 15 of 71 BACK TO MAIN 3M EnvironmentalLaboratory Report No. W1880 In the absence of a clear trend relating the degradation rate to sample pH, it is appropriate to "pool" the data from all pH levels and to determine the degradation rate using the entire data set. Figure 3 illustrates the results of this pooled analysis according to Equation 1, and Table 5 summarizes the results of the analysis. I Table 5. Degradation Rate and Half Life of N-MeFOSE Alcohol in Aqueous Buffered Solutions Using Data Pooled Over pH Levels. Calculated Calculated Observed Rate Percent ( 2 4 Rate Rate Calculated (20) Half Constant at Constant Uncertainty Constant at Half Life Life Range 50" C at 50" C 25" C at 25C at 25" C 1 (day? I 0.0030 I (daY-"1 66 (day-? I I I 0.00030 I (years) 6.3 I (Yea=) I 3.8to 19.4 Figure 2. Observed N-MeFOSE Alcohol Degradation for Various pH Levels 0.10 I - -0.05 \ -0.10 & -0.15 = -0.20 -0.25 1-0.30 -0.35 0 . . I _ pH . . 1.5 pH 3.C I , I , pH 5.C - - - . pH 7.C - - _ - pH 9.0 pH 11 10 20 30 40 50 Time (days) Page 16 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Figure 3. Pooled N-MeFOSE Alcohol Data and Slope Regression. -0.40- Dotted Lines: 20 Uncertainty Limits (slope and intercept) -0.50 I I I I We also monitored the concentration of the hydrolysis product perfluorooctane sulfonate (PFOS), but never observed this compound at levels above its limit of quantification (LOQ, equal to 15.7ng/mL). The initial N-MeFOSE alcohol concentration(473ng/mI) and the PFOS LOQ provide a second estimate of the N-MeFOSE alcohol half-life (see in Appendix B, Equations B32 and B33). The maximum degradation rate is given by Equation 3: k, I (kp),, = -t A F 9 Po A t m=l and the minimum half-life is given by Equation 4 Eq. 3 Eq. 4 Page 17 of 71 __- BACK TO MAIN 3M Environmental Laboratory Report No. W1880 We note that in both Equations3 and 4, the initial N-MeFOSE alcohol concentration (Po) and the PFOS LOQ (A;:$) are molar quantities. Table 6 presents the results of the calculation. ITable 6. At (days) 49 Degradation Rate Solutions and Half Life of N-MeFOSEAlcohol in Aqueous Based on PFOS Limit of Quantification Buffered I Maximum Maximum Calculated [Po 1 A;% Observed Rate at 50" C Calculated Rate at 25" C Half Life at 25" C (nmlml) (nmlml) (day-') (day-' ) (years) 0.85 0.029 7.0x I O 4 7.0x IO-' 127 Page 18 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Conclusions We have performed a study of the aqueous hydrolytic degradation 2-(N-ethylperRuorooctanesu1fonarnido)-ethylalcohol (N-MeFOSE Alcohol). Six different pH levels were included in the study, which1were carried out at 50C and extrapolatedto 25C. Our results based on direct observation of the N-MeFOSE alcohol concentration indicate no clear dependence of the degradation rate of N-MeFOSE alcohol on pH. From the data pooled over the six pH levels, we estimate that the hydrolytic half-life of N-MeFOSE alcohol at 25C lies between 3.8 and 19.4 years, with the most likely value of 6.3 years. The concentration of the compound PFOS, a likely hydrolytic product of N-MeFOSE Alcohol, was monitored throughout the study, but remained undetected above its limit of quantification (LOQ = 15.7 ng/mL). Using the LOQ for PFOS and the initial N-MeFOSE alcohol concentration (473 ng/ml), and assuming PFOS is the only hydrolytic product of N-MeFOSE Alcohol, the data indicate that the hydrolytic half-life of N-MeFOSE alcohol at 25C is greater than or equal to 27 years. Page 19 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 References ` "Fate,Transport and TransformationTest Guidelines: 835.21IO: Hydrolysis as a Function of pH,"U.S. EPA Office of Prevention, Pesticidesand Toxic Substances, publication number 712-C-98-057, January 1998. "ExperimentalPhysical Chemistry",F. Daniels, et al., McGraw Hill Book Co. (New York), p. 1:31, 1962. Page 20 of 71 Signatures ~~~ ~ BACK TO MAIN 3M EnvironmentalLaboratory Report No. W1880 Qilliam K. Reagen, Ph.D., Laboratory Management 0 3/3b/u/ Date Page 21 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Appendix A: Analytical Method - ETS 8-178.0, "Preparationof 2-(NMethylperfluorooctanesulfonamido)EthylAlcohol (N-EtFOSE .Alcohol) Hydrolysis Samples and Analysis by High Performance Liquid Chromatographywith Mass Spectrometry Detection." This Appendix presents the analytical method employed in this study. Page 22 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 3M ENVIRONMENTAL LAaoRATORY METHOD PREPARATIONOF 2 - ( N - M E T H Y L P E R n v o R o O ~ ~ ~ O N A M I D O ) - E T H ALCOHOL (N-MJCFOSE ALCOHOL)HYDROLYSSIASMPLESAND ANALYSISBY HIGH PERFORMANCELIQUIDcEIROMATOGFtAPEY WITH MASSSPECTROMETDREYTECTION Method Number: ETS-8- 178.0 Approved by: Initial Date -_ Adoption Date: qb2/@ Revision Effective Date: &/z. M' Date Page 23 of 71 BACK TO MAIN 3M EnvironmentalLaboratory Report No. W1880 1.0 SCOPE AND APPLICATION 1.1 This procedure defines the steps for analysis of 2-@ImethylperfIuorooctanesulfonamido)-ethyl alcohol (N-MeFOSE alcohol) hydrolysis samplesby high performanceliquid chromatography (HPLC)with mass spectrometry (MS)detectionand quantitation. It is based on EPA OPPTS: 835.2110(Reference 18.1). N-MeFOSE alcohol and the potential hydrolysisproduct perfluomoctanesulfonate0 (PFOSanion) are detected and quantified by thismethod. Internal standards for the method are 2-(N-ethylperfluorooctaaesulfo~mido)ethy~alcohol (N-EtFOSEalcohol) and 3,3,4,4,5,5,6,6,7,7,8,8,8-tridecafluorooctane sulfonicacid anion (THPFOS). This method may also be used to screen for the presence of FOSA (perfluomctanesulfonamide). Representativestructuresare shown in AttachmentA. 1.2 Compatible analytes. 2-(N-methylperfluorooctanesulfonamido)-ethyl alcohol (NMeFOSE alcohol), perfluorooctanesulfonate(PFOSanion), 243ethylperfluomoctanesulfonamide)-ethylalcohol (N-EtFOSE alcohol), 3,3,4,4,5,5,6,6, 7,7,8,8,8-tridecafluorooctanesulfonic acid anion (THFFOS) and perfluorooctanesulfonamide (FOSA). 13 This is a performance-based method. Target analyte or surrogate matrix spike recoveries (100 f 25%) are usedfor each sample matrix to evaluatemethod performance. (Refer to Section 10 for the quality control parameters to be analyzed by this method. Refer to Section 14 for the quality assurance evaluation criteria for this method,) 2.0 SUMMARY OF METHOD 2.1 Aliquotsof N-MeFOSE alcohol stock solution are addedto vials that contain buffers at pH 1.5,3.0,5.0,7,0,9.0 and 11.O. The vials are then placed in an orbital incubator/shakerset at 50.0 f 3 OC. Sets of vials are removed at designatedintervalsand the date and time recorded. The aqueous sample from the hydrolysis of N-MeFOSE alcohol is diluted tenfold with methanol (MeOH). The parent compound,N-MeFOSE alcohol, and the PFOS hydrolysis product are separated on a Dionex IonPacONG1 reversed-phase HPLC column using an ammoniumacetate/MeOHsolvent gradient,with detectiodquantitationby electsospray ionization mass spectrometry in the negative mode. 3.0 DEFINITIONS 3.1 Solvent blank, A sample of a.nalyte&ee medium (for example, methanol) that is not taken through the sample preparation process. This blank is used to evaluate instrument contamination. 3.2 Sample duplicates. Two samples taken from and representative of the same sample source and separately carried through all steps of the extractionand analytical procedures in an identical manner. Duplicate samples are used to assess variance of the total method, including sampling, extraction, and analysis. 3.3 Matrix spike (MS). Prepared by adding a known mass of target analyte to a specified amount of a sample matrix. This assumes that an independent estimate of target analyte concentrationis available. Matrix spikes are used to determinethe effect of the matrix on method recovery efficiency. ETS-8-178.0 Prep. ofN-MeFOSE Alcohol Hydrolysis Samples and Analysis by H P L W Page 24 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 3.4 Calibration standard. A dilutionof various amounts of a stock,intermediateor purchased standardto achieve standard sofutionsin a concentration range of interest. Hydrolytic half-lives resulting from these analyses are calculated based on analytical ratios and not absolutenumbers. Therefore, results do not depend on the purity of the standards used. 3.5 Internal standard. A known amount of a compoundor element similarin analytical behaviorto the compound(s)or element@)of interest, added to all samplesand standards, and carried throughthe entire measurementprocess (post-hydrolysis, after f d dilution). It provides a reference for evaluating and controIlingthe precisionand bias of the applied analytical method. 3.6 Surrogate. An organiccompound which is similarto the target analyte(s) in chemical composition and behavior in the analytical process, but which is not normally found in the sample(s). In hydrolysis studies, surrogate is added to CCVs, samples, sample duplicates, and matrix spike samplesalong with the test d y t e @re-hydrolysis). 3.7 Continuing calibration verification (CCV). Standards analyzed during an analytical runto verify the continued accuracy of the dibration curve. This solution may or may not be prepared h m a different source or lot number than the calibrationcurve standards. 3.8 Dilution. A step in the hydrolysis study procedure in which a solvent is added to the test analytehuffersolutionto prepare it for instrumentalanalysis. This step occurs after the vials are removed fiom incubation and before the samples are analyzed. If the solvent used is misciblewiththe test d y t e h u f f e r solution,the diluting solventis merely added and mixed. If the diluting solventis non-miscible,a liquid-liquid extractionis performed. 3.9 Limit of quantitation (LOQ). The lowest concentrationthat can be reliably measured withinspecifiedb i t s of accuracy (see Sections 14.1 and 14.2) and precision (see Sections 14.3 and 14.4) during routine laboratory operating conditions. The LOQ is generally 5 to 10 times the minimum concentration with a 99% confidence limit that the concentrationis greater than zero. However, it may be nominally chosen within these guidelines to simpIifjldata reporting. For many analytes, the LOQ is selected as the lowest non-zero standard in the calibration curve that is greater than 4 times the level of the solvent blanks. Sample LOQs are highly matrixdependent. 3.10 Accuracy. The closeness of agreement between an experimentally determined value and an acceptedrefmencevalue. When applied to a set of observed values, accuracy is a combinationof a random (precision)and a common systematic(bias) component. For purposes of the study, the acceptance criterion is 75% to 125% of the nominal value. 4.0 WARNXNGS AND CAUTIONS 4.1 Health and safety warnings 4.1.1 Wear the proper lab attire for all parts of this procedure. Wear gloves and proper eyewear at all times. 4.1.2 Handle all solvents in a hood for all parts of the described samplepreparation procedure. Wheneverpossible and practical, dilute sampleswith solvent in a hood. 4.1.3 For potential hazards of each chemical used, refer to material safety data sheets, paclcing materials, and the 3M EnvironmentalLaboratoryChemical Hazard Review. ETS-8-178.0 Prep. of N-MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS Page 25 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 4.2 CaUtiOllS 4.2.1 All glassware in which standardsareprepared shouldbe triple-rinsedwith 1:1 acetone/MeOHto reduce the possibility of contamination. 4.2.2 Ensure that the HPLC mobile phases are prepared prior to beginning a run sequence, and that there is sufficientquantity to completethe run. Do not allow the pump to run dry. 4.2.3 Ensure that before starting the run sequencethere is ample hard disk space on the computer to save all run data 43.4 Ensure that there is enough nitrogen in the supply tankto complete sequence runs. 5.0 INTERFERENCE 5.1 Contaminantsin solvents, reagents, glassware,and other sampleprocessingor analysis hardware may cause interference. Use the routine analysis of laboratory solvent blanks to demonstratethat there is no such interfimnce. 5.2 Contaminationfrom columns, HPLC tubing, and detector componentsmay cause interferenceat low detectionlevels. The routine analysisof solventblanks must be used to demonstratethat there is no such interference. 6.0 EQUIPMENT 6.1 Analytical balance sensitive to 0.1 mg 6.2 Incubator/shaker capableof maintaining temperatureat 50.0 f 3 "C 6.3 Hewlett-Packard(HP) 1100HPLC System, or equivalent 63.1 Pump, binary, Model 01312,or equivalent 63.2 Solvent degasser, Model G1322A or equivalent 63.3 Autosampler,A L S Model 01313A,variable injection volume capable 63.4 Columnheater,Model G1316A 6.4 Dionex IonPacQNGI Guard column, 4x 35 mm, or equivalent 6.5 Mass spectrometer.Hewlett-Packard MSD Model G1946A, or equivalent,operating in electrospray-negativeSIM mode 6.6 Clock, digital.Only one clock should be used,to insureunambiguousdocumentationof the c o m t performanceofprocedures. 6.7 pH meter. Coming Model 308 pWTemperature Meter with 3-in-1 gel-filled combination electrode(pH./reference/temperature),or equivalent 6.8 Reiiigeratorcapableof maintaining4 f 3 "C 6.9 Data system. A personal computer capabIe of controlling the HPLC system as well as recording and processing signals from the detector, Hewlett-Packard ChemStation@ Version A.06.01 or later 7.0 SUPPlJEs AND MATERIALS 7.1 Vials, 40 mL, VOA (I-Chem or equivalent) 7.2 Crimp cap autovials, 1.8 mL 7.3 Labels 7.4 Graduated pipets, glass, disposable, l ' d to 10 mL 7.5 Pasteia pipets, glass, disposable ETS-8-178.0 Prep. of N-MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLC/MS Page 26 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 7.6 Hamilton Gastight@syringes (precisionf1% of total volume), 10 pL-IOOOpL 7.7 Volwnetric flasks,various sizes 7.8 Beakers,glass,various sizes 7.9 Automatic pipettor, capable of dispensing 10-5000 pL 8.0 REAGENTS AND STANDARDS 8.1 Methanol (MeOH). HPLC/SPEC/GC&? fiom EM Science, or equivalent 8.2 Acetone. HPLC/SPEC/GC grade fiom EM Science, or equivalent 8.3 18.0 Mf2 water. Water with lower resistance must not be used. 8.4 Ammonium acetate, 2 mM in water. This solution is chromatographicsolventA (see Section 12.3.1). (Example: An acceptable eluent solution is made by adding 0.15g ammonium acetate crystals to a 1-L volumetric flask containing about 500 mL 18.0 Mi2 water, adding 10 mL of methanol, diluting to the mark with 18.0 M!2 water and mixing.) 8.5 Stock, internal standard and calibration solutions All weights should be recorded to the nearest 0,0001 g in a standardspreparation log: 8.5.1 N-MeFOSE alcoholprepared in acetone. (Example: A stock solution is prepmd at a concentrationof approximately30,000 pg/mL by weighing 0.3 g of NMeFOSE alcohol in a 10-mL volumetric flask and bringing to the mark with acetone. This solution is diluted in MeOH to make additional, appropriate standards.) 8.5.2 N-EtFOSE-alcohol internal standard prepared in acetone. (Example: A stock solution is prepared at a concentration of approximately 30,000 pg/mL by weighing 0.3 g of N-EtFOSE-OH in a 10-mL volumetric flask and bringing to the mark with acetone. This solution is diluted in MeOH to make additional, appropriate standards.) 8.5.3 Perfluormctanesulfonate(PFOS)prepared in acetone. (Example: A stock solution is prepared at a concentrationof approximately3000 pg/mL by weighing 0.06 g of PFOS in a 20-mL volumetric flask and bringing to the mark with acetone. This solution is diluted in MeOH to make additional, appropriate standards.) 8.5.4 3,3,4,4,5,5,6,6,7,7,8,8, 8-t~idecafluorooctanesulfonicacid (THPFOS) internal standard prepared in MeOH. ( Example: A stock solution is prepared at a concentration of approximately20,000 pg/mL, by weighing 0.2 g of THPFOS in a IO-mt volumetric flask and bringingto the mark with MeOH. This solution is diluted in MeOH to make additional, appropriate standards.) 8.6 Buffers for calibrationof pH meter PurchasedpH calibrationstandardsof pH 4.0,7.0, and 10.0 (suppliersvary). 8.7 Bnffer solutions for hydrolysis study. Prepare buffer solutions of pH 1.5,3.0,7.0,9.0 and 11.O using guidelines from CRC Handbook of Chemistryand Physics (Reference 18.2). Prepare buffer solution of pH 5 .O using guidelines fiom Fate, Transport and TransformationTest Guidelines(Refqence 18.2). Preparethe buffer solutionsin I-liter quantities. Calibrate a portable pWtemprame meter using purchased pH calibration stanhds of pH 4.0,7.0, and 10.0, and measure the pH of all buffer solutions. Prepare buffer solutionsofpH 1.5,3.0,5.0,7.0,9.0 and 11.0 at ambient room temperature. The ETS-8-178.0 Prep. of N-MeFOSE Alcohol Hydrolysis Samples and AnaIysis by HPLCMS Page 27 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 concentrationsare given beIow. Record finalpH measurements of all buffers. Store buEers in sealed glass containers. 8.7.1 pH1.5 8.7.1.1 207 mL of 0.1 N HCI (reagent grade) 8.7.1.2 125 mL of 0.2 M KC1 (reagent grade) 8.7.13 Add 18.0Ma water to about 900 mL total volume 8.7.1.4 Adjust pH to 1.5with additional 1 N HCl 8.7.15 Bring to a find volume of 1 L with 18.0MSZ water 8.7.2 pH3.0 8.73.1 223 mL of 0.1 M HCl (reagent grade) 8.7.2.2 500 mL of 0.1 M potassium hydrogen phthalate (reagent grade) 8.7.2.3 Add 18.0 Msz water to about 900 mlL total volume 8.7.2.4 Adjust pH to 3.0 with 1 N HCI or 1 N NaOH 8.7.2.5 3ring to a finalvolumeof 1L with 18.0 MA2 water 8.73 pH5.0 8.73.1 467 mL of 0.1M NaOH (reagent grade) 8.7.3.2 500 mL of 0.1 M monopotassiumcitrate (reagent grade) 8.733 Add 18.0MS2 water to about 900 mL total volume 8.73.4 Adjust to pH of 5.0 with 1N NaOH or 1 N HCI 8.73.5 Bring to a final volume of 1L with 18.0MS2 water 8.7.4 pH7.0 8.7.4.1 500 mL, 0.1 M KH2P0, buffer (reagent grade) 8.7.4.2 291 mL 0.1 N NaOH (reagent grade) 8.7.43 Adjust to pH 7.0 with either 1 N HCl or 1 N NaOH 8.7.4.4 Bring to a final volume of 1 L witk 18.0 M,cz water. 8.7.5 pH9.0 8.7.5.1 500 mL 0.025 M sodium borate decahydrate (reagent grade) 8.7.5.2 46 mL of 0.1 N HCl (reagent grade) 8.7.53 Add 18.0 MSa water to approximately900 mL 8.7.5.4 Adjust to pH 9.0 with either 1 N HCl or 1 N NaOH 8.7.5.5 Bring to a finalvolume of 1 L with 18.0 Mn water. 8.7.6 pH 11.0 8.7.6.1 500 mL 0.05 M NaHCO, (reagent grade) 8.7.63 227 mL 0.1 N NaOH (reagent grade) 8.7.63 Add 18.0Mi2 water to approximately900 mL 8.7.6.4 Adjust pH to 11.0 with IN NaOH 8.7.6.5 Bring to a final volume of 1 L with 18.0I N 2water 8.8 Test d y t e and spike solutions: 8.8.1 N-MeFOSE alcohol test analyte solution with THPFOS surrogate. @xample: An d y t e solutionof N-MeFOSE alcohol,TN-A-1282, at approximately500 pghL and THPFOS at approximately400 p g / d is used (a dilutionin MeOX of the solutionsprepared in Sections8.5.1 and 8.5.4). A 10-pL aliquot of this solutionaddedto 1 mL buffer (stepfound in Section 12.1.5)resultsin a final concentrationof approximately500 n g / d N-MeFOSE alcoholand ETS-8-1.78.0 Prep. of N-MeFOSE Alcohol Hydrolysis Samples and Aualysis by HFLC/MS Page 28 of 71 BACK TO MAIN 3M EnvironmentalLaboratoryReport No.W1880 approximately400 n g / d TypFOS after MeOH dilution (step found in Section 12.1.13)]. 8.8.2 N-EtFOSE alcohol internal standardsolution. pxample: An d y t e solution of N-EtFOSEalcohol, TN-A-1284,at approximately30 pgmL is used (a thousand-foId dilution in MeOH of the solutionprepared in Section8.5.2). A 100-pLaliquot of this solutionaddedto 1mLbuffer (step found in Section 12.1.5) results in a final concentrationof approximately300 n g / d after MeOH dilution (step found in Section 12.1.13)]. 8.83 Spiking solution. [Example: A spiking solution is prepared by adding 10-pL of the N-MeFOSE dcohol stock solution (Section 8.5.1) to a 10-mL volumetric flask and diluting to the mark with methanol. A 70yL aliquot of this solution added to the 1 mL sample (Section 12.1.15) results in a finaIspike concentrationof approximately210 n g / d ofN-MeFOSE alcohol]. 9.0 SAMPLEHANDLING 9.1 Record times of initial preparation and dilution on the fluorochemical degradation (llydrolysis) analysis sample preparation sheet (Attachment B). 9.2 For T i e 0 samples, aliquot only the 1mL of buffer into the vials. DO NOT spike with test d y t e . Storethe vials at mom tempera- until ready to analyze. Then proceed fiom Section 12.1.12. 9.3 Once the 9.0 mL of diluting solvent has been added to the hydrolysis mixtures, the samples should be analyzed as soon as possible. Alternatively, aliquots of the methanoldiluted samples shouldbe refiigeratedat 4 f 3 O C until analysiscan be performed. 10.0 Q u mCONTROL 10.1 Sample Duplicates. Prepare and analyze all samplesin duplicateto provide a measure of the precision of analysis. 10.2 Matrix spikes. Prepare a post-hydrolysis matrix spike sample (Section 8.8.3 and 12.1.12) for each interval and pH level used in the study. Concentrationsof the spike should be approximately equal to a mid-range calibration standard. The matrix spike sampleshouldbe analyzed immediatelyfollowing the sampIe duplicatesto which it corresponds. The analyst shall acceptpercent spikerecoveries of 100f 25%. Spike recoveries outside of this range should be noted. Appropriate steps must be taken to correctthe problem before analysisis allowedtoproceed. Beforethe analysisis allowed to prcceed, consult with the Team Leader or designee for direction and final acceptance or rejection of the analyticai run. 10.3 Solvent blank. Solvent blanks shouldbe runbefore and afterevery calibration w e , CCV:,and after no more than20 injections. Acceptablevalues for the blank are values less than25% of the LOQ standard. If analyte carryover is a problem, use back-to-back solvent blanks. 10.4 ContinuingCalibration Verification (CCV). A standard analyzed periodically during an analyticalrun to verify the continuedaccuracy of the calibrationcurve and is runin tandem with the solvent blank. This solutionmay be prepared froma different source or lot number than the calibration curves standards. ETS-8-178.0 Prep. ofN-MeFOSE Alcohol Hydrolysis Samplesand Analysis bv HopLCMIS Page 29 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 10.5 Internal standard. Internal standardis addedpost-hydrolysis (after finaldilution) to all standards, samples, and matrix spikes at a constant concentration. 11.o CALIBRATIONAND STANDARDIZATION 11.1 Standard preparation. Prepare six calibration standardscontainingN-MeFOSE alcohol,N-EtFOSE alcohol, PFOS and THPFOS in 9:1Me0H:buffer for each pH level. Standardsfiom approximately75 nglmL to 750 n g / d of N-MeFOSE alcoholand 3 ng/mI, to 100ng/mL of PFOS are suggested. A level of approximately400 ng/mL THPFOS surrogateand 300 n g h L of N-EtFOSE alcohol internal standardin each calibration standard is suggested. 11.2 Calibration standards. Analyze the calibration standards at the beginning and end of the run. Average the peak area response from both curves. Use the data reduction software prograrn for linearregressioncalculationsto relate the analytepeak area ratio versus amount ratio, using internal standard calibration. Use N-EtFOSE alcohol as the internal standard for N-MeFOSE alcohol quantification, and THPFOS surrogate as the "internal standard"for PFOS quantitation. Quadraticregression may be used if data review showsthis to be a consistent and more accuraterepresentation of the instrument response. Consultwith the Team Leader for directionprior to performingthe quadratic calibration methodology. 12.0 PROCEDURES 12.1 Sample and spike preparation 12.1.1 Before spikingwith any of the stock standards,transfer approximately 1 mL of the solution to an autovial and cap the vial. Use this smaller volume for spiking to minimize the effects of evaporation from stock solutions and to prevent contamination of the larger volume of stock solution. 12.1.2 Determinethe number of time intervalsthat willbe analyzed. Each interval will have three vials for each pH, multiplied by the number of pHs analyzed. One vial at each level will be labeled as sample, duplicate, and spike. 12.13 Obtainthe appropriatenumber of 4 0 4 VOA vials with caps and cardboard boxes. Prepare appropriate sample preparation worksheets, create labels, and a f k themto the vials. The labels should inchde the samplenumber and i.d., temperature, pH, time interval,test analyte, and date of preparation. Record the pH of each buffer solution. 12.1.4, Removethe cap of the VOA vial and add 1mL of the appropriatebuffer solutionto all of the pre-labeled vials. Always replacethe cap immediately after any addition to minimize evaporation. 12.1.5 Put "Time 0" samples aside at this point. For all other samples, continue on to section 12.1.6. 12.1.6 To all of the vials, add 10 pL of the combined N-MeFOSE alcohol and THPFOS analyte solution(Section 8.8.1) with a 25-pL HamiltonGastight@ 12.1.7 syringe. Make sure that the cap has been firmly tightened and place the samples back in the cardboard case. ETS-8-178.0 Prep. of N-MeFOSE Alcohol Hydrolysis SampIesand Analysis by HPLCMS Page 30 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 12.1.8 Place the case into a pre-warmed incubator/shakerfor the appropriatetime. Record the time, temperame, and rate of shaking. The temperature is determined by the conditionsof the experiment. Continuetomanually monitor the incubator temperature daily during the entire incubation. Record the temperature on the samplep r e p t i o n sheet (AttachmentB). 12.1.9 Store "Time 0" samples at room temperature until the time of analysis. 12.1.10 Remove the case from the incubator at the designated preset time. 12.1.11 Remove the vials from the case and place in racks. Allow the vials to cool for approximately 15 minutes to room temperature. 12.1.12 While vials fiom the first time point are cooling, spike the stored "Time 0" samples with test anaIyte solution (Section 12.1.6). Then continue on to section 12.1.13 with all samples. 12.1.13 Add 9 mL of methanol to each vial. 12.1.14 Using a 100-pLgas tight syringe, add 100pL ofN-EtFOSEalcohol internal standardsolution(Section 8.8.2)to each sample and spike vial. 12.1.15 Using a 100-yL gas tight syringe, add 70 pL of N-MeFOSE alcohol spiking solution(Section 8.8.3)to the Samplespike vials. Shakethe vials for three minutes by hand or Vortex mixer to mix the contents and extract any analytes that may have adsorbed to the vial. 12.1.16 Aliquot approximately 1 mL of each sample to the appropriately labeled autovial, cap, and refrigerate at 4 f 3 OC until analysis. that 12.2 Instrument set up 123.1. Check the appropriate HPLC column is in the instrument for analysis. 12.2.2 Check that the correct eluent solutions are in bottles to be used and that enough is availableto completethe sequencerun, 12.2.3 Place the samples in the autosamplertray and constmct a sequence table with appropriatecalibrationstandards, calibrationcheck standardsand solventblanks. 12.2.4 Verify that alf samplesand standardsare positioned correctly. Enter sequence information (sample or standard ID, method name). Use one injection per sample. 12.2.5 Save sequence as analysis date and instrumentletter (e.g. on March 14,1999,save sequencetable as 0314994. Saveall datato a subdirectorylabeIed with analysis date. (e.g. 031499). 12.2.61 Set post-sequence command macro to shut down system after the run is completed (Example:"STANDBY"on HPllOO/MSD systems). 12.3 HPLC set up: 12.3.1 Analysis of N-MeFOSE alcoholhydrolysis samples in buffersat pH levels 1.5, 3.0,5.0,7.0,9.0 and 11.0. ' Column: Dionex IonPace NG1 Guard column, 4 x 35 mm, or equivalent SolventA: AmmoniumAcetate 2mM in water (with 1% MeOH). Solvent B: Methanol Recommended Sobent Gradient: ETS-8-178.0 Rep. of N-MeFOSE Alcohol Hydrolysis Samples and Analysis by J3PLC/MS Page 31 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 TIME (UIN) 0.0 1.O 4.0 11.0 ~ ._ %A XB FLOWRATE 60 40 0.3 mlfmin 60 40 0.3 mumin 5 95 4.3 mumin '5 95 0.3 mlfmin 12.4 Recommended mass spectrometer set up': MSD: I I I I I Ionizationmode API-ES ~ Polarity Negative Acquisitlon mode SIM SfM resolution High Time filter Enabled I Peakwidth I 0.25 min I Gain 1.o Fraamentor 70 I owell time I 183msec I Capillary voltage 3500 Drying gas Nitrogen Nebulizer pressure 30 psig Drying gas flow 8 Umin Drying gas temp 300' C 12.5 Auto-sampler setup': AUTOSAMPLER: -AUTO-SAMPLER PROGRAM: -INJECTIONVOLUME: ALS Model G1313A None 5.0 pL 12.6 Ions used for identification and quantification: APPROX. RETENTION TIME (MIN) 8.4 8.6 6.2 COMPONENNTAME MeFOSE-OH EtFOSE-OH PFOS DESCRIPTION QUANTIFICATION ION K)H1 MONITOR Test W y t e 616 Internal Standard 630 Potential Degradation 499 Product 500 I Page 32 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 5.9 THPFOS Sumgate * FOSA Qualitativeuse onIy Wo calibrationstandards maiyzCd, so no retentiontimes available. 427 None ~ __ None 498 12.7 Sample analysis 12.7.1 Enter the standard,sample, and QC informationinto the sequencetable. Analyze calibrationstandardsfirst,then up to 20 injections,followed by the calibration standards. If more than 20 injections are to be run,analyze a continuing calibrationstandard (CCV)after every 20 injections and runthe calibration standards again at the end of the sequence. Run solvent (or method) blanks after the highestcalibrationstandard,before and afterthe CCV, and after the set of samples to check for any analyte canyover. 12.7.2 Place standards,samples, and QC (matrixspikes, sampleduplicates,and blanks) into the autosamplertray accordingto the order they are listed in the sequence. 12.73 Identifythe electronic acquisitionfiles with an appropriateprefix (eg. MeFOS). Do not exceedfive charactersif the sequencecontainsmore than 99 lines. 12.7.4 Save sequence as analysis date (e.g. on March 14, 1999, save sequence table as 031499s). Save all datato a subdirectorylabeled withanalysisdate (e.g. 03 1499). 12.7.5 Start the sequence. 13.0 DATAANALYSIS AND CALCULATIONS 13.1 Peak Evaluation. Peaks must be symmetricin shape and identifiedby extracting compound-specific ions. Peaks considered for calibration must have peak heights greater than 5 (five) times the baseline noise for that region of the chromatogram. Peak area integration is from baseline to baseline using automatic or manual integration. 13.2 Calculation of Rate Constant (k). Calculatethe test analyte concentrationsin each of the pH matrices using the curves obtained from the calibrations. Assuming first-order - - (-e. kinetics a rate constant (k)can be determined by plotting: [N MeFOSE OH] [N- MeFOSE -OH& versus minus elapsedtime The subscripts t and 0 refer to analyte concentrationsdeterminedat some elapsedtime t and at t = 0, respectively. The slope of the resulting line is k. The ? value for this plot should be > 0.80. For ? values less than this,consultthe Team Leader or designee. 133 Target analyte concentrations. Calculatethe Me-FOSE alcohol and PFOS concentrationsin each of the pH matricesusing the curves obtained from the calibrations. 13.4 Matrix spikes. Calculatethe percent recovery for each of the matrix spikes. Calculate the mtrk spike percent recoveries using the following equation: % Retmvery=[observed soiked samde result -observed samde result) x 100 Actual amount spiked Using the observed matrix spike recoveries, calculate the average spike recovery. ET'S-8-178.0 Prep. of N-MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLCIMS Page 33 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 13.5 Sample Duplicates. Calculate the relative percent deviation (RPD) for the duplicate samples: WD= IA-SI ~ 1 0 0 % (A+B)/2 Where A- the concentration measured in the sample B= the concentrationmeasured in the sampleduplicate 14.0 METHOPDERFORMANCE 14.1 Accuracy. For purposes of the study, the acceptancecriterion is 75% to 125% of the nominal value. 14.2 Coefficient of determination (13. The coefficient of determination (3) for the calibration curves should be 0.99 or greater. The curves should be examined closely for linearity and intercept, particularly for accuracy of quantitation at the low and high ends of the curve. The accuracy of all standardsused for calibrationmust be within75-125%. On occasion it may be necessary to use exponential or quadratic fits of the data, usually when broad range curves (greater than 3 orders of magnitude between the low and high concentrationstandards)are used. Document in the raw data the technicaljustification for using quadratic equations. 14.3 Matrix spikes. The analystshalI acceptpercent spike recovery values of 100f 25%. Spikerecoveriesoutside of thisrange should be noted. Consultwith the Team Leader or designeefor direction, and for final acceptanceor rejection of the data. Data that are used in finalreportthat isdeemed out of control willbe required to have a technical justification for why the data are being used,documented in the final reportand raw data. 14.4 Sample duplicates. The analyst shall accept %RPD (See Section 13.5)values e 25%. %RPD values > 25% should be noted. RPD values of 25% or greater should be noted. Appropriate steps must be taken to correct the problem before analysis is allowed to proceed (e.g. sample re-runs,additional blanks, etc.). Consultwith the Team Leader or designeefor direction, and for final acceptanceor rejection of the data. 14.5 Continuing calibrationverification (CCV). Ifthe percent difference for the amount of quauthted analyte is greater than25% &omthe true value relativeto the initial standard curve, stop the run. Only those samples analyzed before the last acceptabIe calibration check standardwill be used. Consultwith the Team Leader or designee for direction, and for final acceptance or rejection of the data. 14.6 Internal standard and Surrogate. Review of the internal standardand surrogatecan be performedby averaging the area response throughoutthe analytical runand calculating the relative standard deviation (%RSD). %RSD values >lo% should be noted. Inconsistenciesin the internal standardpeak area may indicate instrumental changes over time. Inconsistenciesin the surrogatepeak area may indicate instrumentalchanges, changes in the test-system, or hydrolysis of the surrogate over time. Consult with the Team Leader or designee for direction and final acceptanceor rejectionof the analytical nul. ,. Em-8-178.0 Prep.ofN-MeFOSEAlcohol Hydrolysis Samples and Analysis by H P L W Page 34 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 14.7 14.8 14.9 14.10 Limit of Quantitation(LOQ).The LOQ is equal to the lowest standard in the calibration curve that is greater than4 times the level of the solvent blanks. If blank contaminationis present, it may necessaryto deletethe lowest standard of the calibration curve. Solventblanks. Solventblanks should show no more thana 5% canyover from a high standard or calibrationcheck standard. If so, two solvent blanks may be necessaryto rule out instrumentalcontamination. If peaks with greater than25% of the peak area of a low standard value are observed in sequential solvent blanks, it is indicative of instrument contamination. The instrument shall be serviced by thoroughly cleaning the electrospray source, and replacing/cleaningcolumns,tubing, etc. Specificity. Analyte specificityis demonstratedby acceptable post-hydrolysisamiyte spike recoveries. System Suitability. Without performing a method validation, system suitabilitycan be demonstratedby acceptable instrumentalchecks (e.g. abbreviated m/z check-tune, or full auto-tune routines). Consult the appropriate instrumental manuals (Reference 18.3). 15.0 POLLUTION PREVENTION AND WASTE MANAGEMENT 15.1 Dispose of sample waste by placing in high or low BTU containers as appropriate. Use broken glass containersto dispose of glass pipettes. 15.2 Collect HPLC solvent waste in the satellite accumulation can. Empty into the flammable storage drum in the hazardous waste collection area on the 2nd floor. 15.3 Use smallerbore columns when possible to minimizewaste generation. 16.0 RECORDS 16.1 Print hard copies of all graphics and data analysis summaries for archiving- . 16.2 Sign and dag all data packages and label wi& instrument ID. 16.3 Fill out the hydrolysissampIe preparation worksheetcompletely, making sureto include all initials and dates. 16.4 Print out the sample sequence table, reduce the size with photocopying and tape the photocopy into the instrument log. Keep the originalcopy for the raw data filespackage. 16.5 Print chromatogramsand quantificationreports for a l l analyses. 16.6 Print calibrationtables and curve informationand store in the raw data file. 16.7 Store hydrolysis sample preparation worksheets in the raw data file. 16.8 Enter all standardpreparation informationin the standardspreparation logbook. Make a photocopy of the logbook page and include the copy in the raw data file. 16.9 Archive electronic data to appropriate media when necessary. 17.0 ATTACHMENTS 17.1 Attachment A. Representative chemical structures 17.2 Attachment B. Hydrolysis sample logsheet ETS-8-178.0 Prep. of N-MeFOSE Alcohol Hydrolysis Samples and Analysis by HPLCiMS Page 35 of 71 BACK TO MAIN 3M EnvironmentalLaboratory Report No. W1880 18.0 REFERF,NCES 18.1 Fate,Transport and TransformationTest Guidelines Officeof Prevention, Pesticides and Toxic Substances(OPPTS) 835.2110 Hydrolysis as a Function of pH, EPA 712-C-98- 057, January 1998. 18.2 CRC Handbook of Chemistry and Physics, 1st Student Edition, "BufferSolutions Operational Definitions of pH," Robert C. Weast, Ph.D., 1988, p. D-87. 183 HP 1100 SeriesLCMSD Reference Collection,Rev. A.OO.O1 June 1997 CD/ROM 19.0 AFFECTED DOCUMENTS 19.1 None. 20.0 FIEVISIONS Revision number Reason for revision Date of Revision ETS-8-178.0 hep.of N-MeFOSEAlcohol HydrolysisSamplesand Analysisby HpLclMS Page 36 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Attachment A. Representative Chemical Structures +\ I I I I I I I Ho N-MeFOSB Alcohol FW = 557.23 I I I I 3+v no N-EtFOSE A l ~ ~ hFoWl = 571.25 FOSA FW499.14 C, Olefin FW-382.07 PFOS FW=500.13 ' C, Hydride FW420.07 . .. . THPFOSFW428.17 . .. Attachment A ETS-8-178.0 Prep. of MeFOSE Alcohol Hydrolysis Samples and Analysis by H[pLC/MS Page 37 of 71 BACK TO MAIN mi: E FF c ~ ~ ~~ BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Appendix B: Kinetics Model This Appendix includes a mathematicaldescription of the kinetics model employed in the study. Page 39 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Kinetics Model B1. Reaction Components and Rates The arguments below are based on the following idealized set of reactions representing the hydrolysisof a parent compound P and its hydrolysis products A,, which number N. The actual hydrolysis reactions that occur under neutral, acidic, and basic conditions are subsumed in these equations, and are assumed to proceedwith pseudo-first order rates k,, (for the parent) and k,, (for the parent's hydrolysis products). P + H,O kP, e n, A, +Yd (m=ltoN) kh A, +H,O a Ym2 (m = 1 to N) where the general symbolsYm1and Ym2represent all the other hydrolysis products. 82. Parent Compound Concentrations Equation B1 indicatesthat the pseudo-firstorder differential change in the parent concentration P is given by which is equivalent to the separable differential equation Equation B4 may be directly integratedto obtain the general solution With the initial condition P(t = 0) = Po,the specific solution to Equation B4 is (-2 P = Po exp n, k,, t l - Po e-kp ( m=l I using the additional definition of the total parent hydrolysis rate Page 40 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 m=l Equation B6 can be re-written in a form that allows a least-squaresestimate of the total parent hydrolysis rate: kpt=-h [t] Using the initial(t =0) measuredvalue of the parent concentration Poand later values P measured at later times t , one can calculate and plot the (linear) quantity c p [- In (P/P,)] versus time and obtain a least -squares estimate of the slope of the line. The resulting slope is the least-squares estimate of the total parent hydrolysis rate. Equation B6 indicates that over a period of time TI/: (the parent hydrolysis half-life) the parent concentration P is reduced through hydrolysis by a factor of two, where A least squares estimate of the parent hydrolysis half-life is therefore available from B3. Product Compound Concentrations The pseudo-first order differential changes in the product concentrations&, (using Equations B2 and B6) are dAm= (n,kpmP- kAmAm)d=t ( nmkpmPeO-kp - kAmAm)dt and the (first order, non-separable) differentialequation governingthe product concentrations is -`A+m k,A, = n,kp,Po e-kpt. dt The "standard form" of Equation B12 is Page 41 of 71 - BACK TO MAIN 3M Environmental Laboratory Report No. W1880 A,,' + S (t) A, = Q(t) where the "function"S(t)is actually a constant: and Q(t) =n,k,,Po e-kpt . The general solution A, to Equation 812 is contained in where and jQ(t)e]S(t')dtd`t + C = n,kp,Poj ekht e-kptdt+ C There are two cases of Equation B18 to consider. In the circumstance that kAm= k, , which occurs only when the hydrolysis rate of the mthproduct is identical to the total parent hydrolysis rate, the general solution to Equation B18 is (for k, = k,) A, + ekpt= nmkpmPtO C and, using the initialconditionA,(t = 0 )= A, , the specific solution to Equation18 is (for k, = k,) A, = (nmkPmPtO+ A r n o ) e - k p. t We note that when k, = k, = 0 (that is, when both the parent and potential product are hydrolytically stable), Equation B7 requires (also) that k,, =0, so Equation 820 becomes Page 42 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 indicating, as required, that the product concentration does not change with time. The circumstance kA, = k, is highly improbable, and is neglected in the remainder of this discussion. However, the reader should bear in mind that the expressions derived below do not hold when the parent hydrolysis rate k, and the product hydrolysis ratek, approach each other. In the more probable case, for which kA, f k, (Le. that the hydrolysis rate of the mth product is different from the total parent hydrolysis rate), the general solution to Equation B18 is and the specific solution to Equation B18 with the initial condition A,(t = 0) = A,, is e-k,t - nmkPrnPO e - k p t . k P - kArn Of greatest interest here is the case in which the product compounds are known to be hydrolytically stable, that is, when k, = 0 for all m. In this case, Equation B23 becomes (for hydrolytically stable products) + A, n,k P Pm o ( l-eelpf) . kP 84. RelationshipsBetweenthe Parent and Compound Concentrations Equations B7 and 824 can be combined to obtain (for hydrolytically stable products) N N k p = n, k,, = rn =1 Page 43 of 71 so that or BACK TO MAIN 3M Environmental Laboratory Report No. W1880 (for hydrolytically stable products) (for hydrolytically stable products) If the changes in the productconcentrationsare all small compared to the original parent concentration,that is, if we may use the expression (valid for -1 I X I 1 ) h ( l + X ) = X - -1 x2+ -1 x 3--1x4 +..... 2 3 4 and Equation B23 becomes (for hydrolytically stable products and IFA, -Amo << Po) k,t N z-[-ZAm ioAmo] Page 44 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 or (for hydrolytically stable products and cm A, -A,, << Po) B5. Parent Half-Life Estimates Based on Limits of Quantification of the Products In every experimentaldeterminationof k, ,there is some set of values A r Q(the "limits of quantitation") below which the product concentrations A, cannot be reliably measured. If during an experiment carried out over the period of timeA t all the product concentrations A, remain below their limits of quantitation, then the maximum possible value of the rate k, is obtained by assuming (for all the products)that 1) A,, = 0 and 2) at time t = A t , the product concentrations have increasedto the values A, = A P Q . With these assumptions, the experimental data indicate that the reaction rate k, is less than some maximum value (kp)- as follows: (for hydrolytically stable products at concentrations below the limits of quantitation) k, I(k,),, = - c 1A k oNQ . Po A t m=l Under the same circumstances and assumptions, the experimental data indicatethat the parent half-life TI': (see Equation 69) is greater than the value (Tyi) . as follows: mm (for hydrolytically stable products at concentrations below the limits of quantitation) The reader should note that Equations 932 and 933 are valid only when both 1) the products are hydrolyticallystable and 2) the concentrationsof all the potential products are measured. Otherwise, the quantity (kp)- in Equation 932 may not actually representthe maximum possible value of the rate constant k p, and the related result in Equation 933 for (Tv;) . is also questionable. mmn Page 45 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 96. Parent Half-Life Estimates Based on Limits of Quantification and Experimental Precision of Product Concentrations In certain experiments, some hydrolysis products are present at quantifiable but essentially constant concentrations over the time ( A t ) of the experiment. In this case, it is the experimental precision of the measured product concentrations, rather than the limits of quantitation, which contribute to the estimate of the maximum value of the parent hydrolysis rate k, . If the set of concentrationsmeasuredfor the mthproduct have the mean value p, and standard deviation o m ,the data do not exclude the possibilitythat the product concentration increasedfrom the initial value 0, -p, to the value 0, + p, at time t = A t . Taking this possibility to be the actual case for the measured products, the maximumvalue of the quantity (A, - A m o )is 2 0 , . This reasoning suggests that the following estimate of the maximum parent hydrolysis rate is appropriate: (for hydrolytically stable products at either 1) constant measured concentrations with standarddeviation Om or 2) concentrations below the limits of quantitation) r 1 v i ) Under these circumstances and assumptions, the experimental data indicatethat the parent half-life TI': is greater than the value (T . as follows: rmn (for hydrolytically stable products at either 1) constant measured concentrations with standarddeviation Om or 2) concentrationsbelow the limits of quantitation) r 3-1 The reader should note that Equations B34 and 835 are valid only when both 1) the products are hydrolytically stable and 2) the concentrationsof all the potential products are measured. 96. Parent Half-Life Estimates Based on the Experimental Precision of Parent Concentrations In certain experiments, the hydrolytic parent remains at an essentially constant concentration over the time ( A t ) of the experiment. In this case, it is the experimental precision of the measured parent concentrationsthat determinesthe maximumvalue of the parent hydrolysis rate k, . If the set of concentrationsmeasuredfor the parent have the mean value p, and standard deviation oP,the data do not exclude the possibility Page 46 of 71 ~ ~~~~~ BACK TO MAIN 3M Environmental Laboratory Report No. W1880 that the product concentration increasedfrom the initialvalue pp - opto the value pp+opat 1:imet = A t . This reasoning suggests that the following estimate of the maximum parent hydrolysis rate is appropriate: (for essentially constant parent concentrations with mean value ppand standard deviationCYp ) kp 5 (kp), = -, 20, ' Pp At Under these circumstances and assumptions, the experimental data indicate that the parent half-life TI/; is greater than the value (T-.)v: as follows: (for essentially constant parent concentrations with mean value ppand standard deviation op) B8. Temperature Dependenceof the Reaction Rate and Half-Life In order to increasethe speed of the reactions of interest, we conducted this experimental study using samples maintained at the temperature 50C = 323 K. Of greater interest are the corresponding results for the environmentally important temperature 25C = 298 K. When the Arrhenius activation energy for a reaction is AHa, Equation 838 " provides the following relationship betweenthe hydrolysisrates (kl and k2)for that reaction at two different absolute temperatures (T,and T,): where R = II .99x I O " Kcal mole-' K-' is the ideal gas constant. Usingthe valueB2 AHa=I8Kcal/mole,the rate ratio kl/k2 at the correspondingtemperatures TI=298K and T2=32?K; is [II- ]] = exp{ 18 = exp(-2.35) = 0.095 (839) k2 1 . 9 9 ~ 1 0 "323 298 Page 47 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Equation 839 indicates that the hydrolysis reactions of interest proceed approximately ten times more slowly at 25C than at the chosen experimentaltemperature of 50C. Accordingly, the rate reactions reported here for the temperature 25C are ten times lower than those measured at 50C, and the hydrolysis half-life estimates reported here for 25C samples are ten times longer than those calculated from the 50C experimental data. References to Appendix B: " I. N Levine, "Physical Chemistry," McGraw-Hill (New York), pp. 498-501 (1978). ''F. Daniels, et al., "Experimental PhysicalChemistry", McGraw Hill (New York), p.131 (1962). Page 48 of 71 BACK TO MAIN 3M Environmental LaboratoryReport No. W1880 Appendix C: Selected Analytical and Kinetics Results This Appendix includes selected sample data and their related kinetics results. Page 49 of 71 BACK TO MAIN 0 Z 5 22i- i- 3 a 3 a I- 3 0 0 w>- If s4 If v) a v) z 0 cj bv) .w Q I- 2 a t BACK TO MAIN r IC c 0 r m s D N BACK TO MAIN I- I- 3 a 3 a I- I- 3 3 0 0 2. 2 !Y I5 fI cn cn -W0 -0 BACK TO MAIN 0,aN-m *C-fgg 0 0 0 C 0 0 0 0 0 0 0 0 0 0 0 0 0 2: 00 000000000000000 000 000 000 000 22 qymr *cf:: BACK TO MAIN z 0 BACK TO MAIN z 0 BACK TO MAIN z 0 n nnnnnn nn nnnnnnnnnnonnnn n o nnnnnnnnn o n n n o n n n BACK TO MAIN 0 a3 h 0 Z we, 99 22 Low ff 88 88 ff if $5 mm sI sS BACK TO MAIN 0 Q) 0 Z BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Pooled N-MeFOSE Alcohol Data and Slope Regression - - - - . . _ _ _ 0.05 ............ - - - - - - - - - - - - - - - - - - - - - - - - - - - - - I 0.00 0 ..... . . . ................... 0 .... .................................................................... . .................................................. : I 0 ---------A -0.05 - '. - -. -0.10 p" - - \ . L -0.15 - -- . c -0.2c - -. -0.25 0 *. * - . 0 0 =-.*O 0 0 Solid Line: y = -3.00E-03~ 1.25E-02 R2 = 1.93E-01 0 --*.* 0 - . . p - - a . *. 0 0 -0.30 -0.35 0 10 20 30 40 50 time (days) Solid Line: Dolted Lines: 2s Uncertainty Limits (slope and intercept) SUMMARY OUTPUT Regression Stabstm Muitiple R 0.4388236411 R Square 0.192566108 Adjusted R Sq 0.17131793 Standard E m 0.09318559 Observations 40 ANOVA Regression Residual Total df SS 1 0.078696282 :1 0.329975058 :39 0.408671341 Intercept X Variable 1 Coeffidents StandardE m r -0.0125162.55 0.029261069 -0.0029963137 0.000995309 %2 s slope uncertainty 66.4 MS 0.078696282 0.008683554 F Significance F 9.062681099 0.004617044 t Stat -0.427744267 -3.010428723 P-value 0.671251755 0.004617044 Lower 95% -0.071752194 -0.005011205 Upper 95% 0.046719635 -0.000981409 Lower 95.0% -0.071752194 -0.005011205 Upper 95.0% 0.046719685 -0.OC0981409 Page 59 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Appendix D: Selected Chromatograms A representative set of chromatograms from the present study is included in this Appendix. Page 60 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Calib. Data Modified : Wednesday, December 29, 1999 9:20:34 ?m Calculate Based on Rel. Reference Window : Abs. Reference Window : Rel. Non-ref. Window : Abs. Non-ref. Window : Internal Standard Peak Area 10.000 % 0.000 min 10.000 % 0.000 min -y M l $ sfl/& \w . weMm' 1s 8 Copy OF fimn)flL MultiplierDilution Sample Amount 1.0000 1.0000 0.00000 p M m 13 3 M w m %J5 P ~ J ~ ~ ~ . Uncalibrat.ed Peaks not reported Partial Calibration : Correct Al.1 Ret. Times: Yes, identified peaks are recalibrated No, only for identified peaks Curve Type - Origin - Weight Linear Included Equal Sj@& sa: SJ.0 cl/fl& ' - - Recalibration Settings: - Average Response Average Retention Time: @L*n'B& Average all calibrations SWY)p& Floating Average New 75%- / 3 L / p L ) L m - Calibration Report Options : MAyWSC Printout of recalibrations within a sequence: Calibration Table after Recalibration Normal Report after Recalibration If tho sequence is done with bracketing: Results of first cycle (ending previous bracket) Sample ISTD Information: ISTD ISTD Amount Name , - - -#- I - - - -[-P-P-b-l- - - - - I - - - - - - - - - - - - - - - - - - - - - - - - - 1 4Q2.60000 THPFOS 2 310.80000 EtFOSE-OH Signal 1: MSDl 427, EIC=426.7:427.7 Signal 2: MSDl 499, EIC=498.7:499.7 Signal 3: MSDl 630, EIC=629.7:630.7 Signal 4: MSDl 616, EIC=615.7:616.7 RetTime Lvl Amount Area Amt/Area Ref Grp Name - -[-rn-i-n-]- S 1 i -- g 1 - - 1 ------[--P--PI--b--l------l----------l---l--l--------------- 5.870 1. 1 402.60000 4.13061e5 9.74674e-4 I1 THPFOS ' 2 402.60000 4.15745e5 9.68383e-4 3 402.60000 4.14516e5 9.71252e-4 4 402.60000 4.16734e5 9.66084e-4 5 402.60000 4.14375e5 9.71584e-4 6 402.60000 4.18593e5 9.61793e-4 11 402.60000 4.07992e5 9.86784e-4 Page 61 of 71 BACK TO MAIN 3M EnvironmentalLaboratory Report No. W1880 Method C:\HPCHEM\l\METHODS\1227-7OI.M RetTime Lvl Amount Area Amt /Area Ref Grp Name - -[r-n-i-n-l - Sig I -- I - - I _ _ [_P_P_b l _ _ _ _ _ I _ _ _ _ _ _ - - _ _ I _ _ _ _ _ _ _-_-_- _ - - I - - - - - _ _ _ _ - _ _ _ _ _ 12 402.60000 4.07169e5 9.88780e-4 13 402.60000 4.09832e5 ,9.82353e-4 14 402.60000 4.09148e5 9.83996e-4 15 402.60000 4.15661e5 9.68579e-4 16 402.60000 4.16714e5 9.66131e-4 6.162 2 1 3.13000 2.19264e4 1.42750e-4 1 PFOS 11 3.13000 1.74464e4 1.79406e-4 2 7.83000 3.32915e4 2.35195e-4 12 7.83000 2.99914e4 2.61075e-4 3 15.65000 5.41038e4 2.89259e-4 13 15.65000 5.80162e4 2.69752e-4 4 31.30000 1.20587e5 2.59563e-4 14 31.30000 1.12160e5 2.79065e-4 5 62.60000 1.97863e5 3.16381e-4 15 62.60000 1.90397e5 3.28787e-4 6 93.90000 2.83971e5 3.30667e-4 16 8.306 4 1 93.90000 2.72629e5 3.44424e-4 78.90000 1.63904e5 4.81379e-4 12 MeFOSE - OH 11 78.90000 1.62533e5 4.85439e-4 2 157.80000 3.53900e5 4.45889e-4 12 157.80000 3.38524e5 4.66141e-4 3 315.60000 6.72851e5 4.69049e-4 13 315.60000 6.51730e5 4.84249e-4 4 473.40000 1.04325e6 4.53774e-4 14 473.40000 1.01321e6 4.67226e-4 6 789.00000 1.67835e6 4.70106e-4 16 789.00000 1.63482e6 4.82621e-4 8.535 3 1 310.80000 7.22548e5 4.30145e-4 12 Et FOSE- OH 2 310.80000 7.26841e5 4.27604e-4 3 310.80000 7.21094e5 4.31012e-4 4 310.80000 7.21008e5 4.31063e-4 5 310.80000 7.22268e5 4.30311e-4 6 310.80000 7.24772e5 4.28824e-4 11 310.80000 7.07642e5 4.39205e-4 12 310.80000 7.00669e5 4.43576e-4 13 310.80000 7.05419e5 4.40589e-4 14 310.80000 7.09949e5 4.37778e-4 15 310.80000 7.18262e5 4.32711e-4 16 310.80000 6.99029e5 4.44109e-4 Page 62 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Batch Run # 1 of 50 Data File C:\HPCHEM\1\DATA\lOO699\MeFOSO5l.D Sample Name: MeOH Blank ..................................................................... Injection Date : 10/7/99 5:09:30 AM Seq. Line : 51 Sample Name : MeOH Blank Vial : 91 Acq. Operator : MTM Inj : 1 Acq. Method : C:\HPCHEM\~\METHODS\FOSESIM.M Last changed : 10/6/99 1:56:00 PM by MTM Analysis Method : C:\HPCHEM\l\METHODS\1227-7OI.M Last changed : 12/29/99 9:23:15 AM by MTM (modified after loading) (Results are from a previously s SIM Analysis (ES-) f o r Et-FOSE-OH,MeFOSE-OH, THPFOS, and PFOS using 4mmx35mm Clionex IonPac NG1 column, S/N 12879. YTM Msm4 ~~ I.G- -.~ L-.fI..pJ . - t s Negauve --. 400 > 5. . . 6""$""8 W1-b NWUW " " Q ' " ' 1250 1000 750 500 S - ] M 6. . " . " 6 " " ~ " " 8"" NegaUVa 9 " " $0 7 $0 m Sorted By Signal CaLib. Data Modified : 12/29/99 9:23:15 AM Multiplier 1.0000 Dilution 1.0000 Sample ISTD Information: ISTD ISTD Amount Name ----I#-------------I--[--~---p---b---l------------ 1 402.60000 THPFOS 2 310.80000 EtFOSE-OH Page63of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Method C:\HPCHEM\l\METHODS\1227-7OI.M I O.4 / .................................. THPFOS at exp. RT: 5.870 MSDl 427, EIC=426.7:427.7 Correlation: 1.00000 Residual Std. Dev.: Formula: y = mx + b 0.00000 m: 1.00000 b: 0.00000 x: Amount Ratio y: Area Ratio PFOS at exp. RT: 6.162 MSDl 499, EIC=498.7:499.7 Correlation: 0.99654 Residual Std. Dev.: Formula: y = mx + b 0.02056 m: 2.79093 b: 2.92806e-2 x: Amount Ratio y: Area Ratio MeFOSE-OH at exp. RT: 8.306 MSDl 616, EIC=615.7:616.7 Correlation: 0.99973 Residual Std. Dev.: Formula: y = m x + b 0.01989 m: 9.19192e-1 b: 6.85505e-3 x: Amount R a t i o y: Area Ratio 0.2 4 I 4 /' EtFOSE-OH at exp: RT: 8.535 MSDl 630, EIC=629.7:630.7 Correlation: 1.00000 Residual Std. D e v . . : Formula: y = mx + b m: 1.00000 0.00000 I b: 0.00000 x: Amount Ratio y: Area Ratio Page 64 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Batch Run # 1 of 50 Data File C:\HPCHEM\l\DATA\lOO699\MeFOS05l.D Sample Name: MeOH Blank Signal 1: M S D l 427, EIC=426.7:427.7 RetTime Type - -[-m-i-nl- - I - - - - - - 5.870 Tota1s wit:hout ISTD(s) : G r p Name --I------------------ . THPFOS 0.00000 Signal 2: M S D l 499, EIC=498.7:499.7 RetTime Type [minl . Area Amt /Area Amount Totals without ISTD(s) : 0.00000 Totals without ISTD(s1 : 0.40000 Totals without ISTD(e) : 0.00000 2 Warnings or Errors : Warning : ISTD compound(8) not found Warning : Negative results set to zero (cal. curve intercept), (PFOS) Page 65 of 71 BACK TO MAIN 3M EnvironmentalLaboratory Report No. W1880 Batch Run # 11 of 5 0 Data File C:\HPCHEM\1\DATA\lOO699\MeFOSO6l.D Sample Name: MFA-010 =ppp=5=e==pD==pp------- ----- --=----------------------------==---------------------- - - - - - - - - H P P C I P I I = I = E - Injection Date : 1 0 / 7 / 9 9 8:11:41 AM Seq. Line : 61 Sample Name : MFA-010 Vial : 51 Acq. Operator : MTM Inj : 1 Acq'. Method : C:\HPCHEM\~\METHODS\FOSESIM.M Last changed : 10/6/99 1:56:00 PM by MTM Analysis Method : C:\HPCHEM\l\METHODS\1227_70I.M Last changed ' : 12/29/99 9:23:15 AM by MTM (modified after loading) (Results are from a previously s SIM Analysis (ES-) for Et-FOSE-OH,MeFOSE-OH, WPFOS, and PFOS using 4mmx35m Dionex IonPac NG1 column, S/N 12879. MTM Sorted By Calib. Data Modified : Mu1tiplie:r Dilution Signal . 12/29/99 9:23:15 AM 1.0000 1.0000 Sample ISTD Information: ~ S T D I s m Amount Name - - -#- I - - - - - -[-P-P-b-l- - - ] - - - - - - - - - - - - - - - - - - - - - - - - - - 1 40:2.60000 THPFOS 2 310.80000 EtFOSE-OH Page 66 of 71 Batch Run Data Fi1e # C 11 :\H of PCH 50 EM\l\DATA\lO 0 6 99\M&FOS 0 6 1 . D BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Sample Name: MFA-010 Totals without ISTD(s) : 0 .ooooo Signal 2: MSDl 499, EIC=498.7:499.7 RetTime Type - -[-m-i-nl- - I - - - - - - 6.203 PBA Area ----------I---------- 1.37520e4 Amt/Area ratio 2.91660e-2 Totals without ISTD(s) : 3.74287e-1 Signal 3: MSDl 630, EIC=629.7:630.7 RetTime Type [mini , Area Amt /Area ratio , Totals without ISTD(s) : 0.00000 Signal 4: MSDl 616, EIC=615.7:616.7 Page 67 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Batch Run # 12 of 50 Data File C:\HPCHEM\1\DATA\lOo699\MeFOSO62.D Sample Name: MFA-011 - - --3======- - . -- - = -P I I I I 3 = = = = I = P P I = = = = = E = E P 1 1 5 = = = = = = E E = = = = = = = ~ a = ~ = = = = = = = = = = Injection Date : 10/7/99 8:29:54 AM Seq. Line : 62 Sample Name : MFA-011 Vial : 52 Acq. Operator : MTM Inj : 1 Acq. Method : C:\HPCHF,M\~\METHODS\FOSESIM.M p#&D ALnaasltyscihsanMgeetdhod :: C1:0\/H6P/C9H9EM1\:l5\6M:E00THOPDMS\b1y22M7T-M7OI.M PMlIrn M Last changed : 12/29/99 9:23:15 AM by MTM / T/+ (modified after loading) (Results are from a previously 8 SIM Analysis (ES-) f o r Et-FOSE-OH,MeFOSE-OH, THPFOS, and PFOS using 4mmx35mm Dionex IonPac NG1 column, S/N 12879. Sorted By Signal Calib. Dat.a Modified : , 12/29/99 9:23:15 AM Multiplier- 1.0000 Dilution 1.0000 Sample IS'IP Information: ISTD ISTLI Amount Name - -#- - I - - - -b-p-b-l- - - - - - I - - - - - - - - - - - - - - - - - - - - - - - - - 1 402:.60000 THPFOS 2 31O.80000 EtFOSE-OH Page 68 of 71 BACK TO MAIN 3M Environmental Laboratory Report No. W1880 Batch Run # 13 of 50 Data File C:\HPCHEM\l\DATA\lO0699\MeFOSO63.D Sample Name: MFA-012 =5II=I====DI=IIPI======31t-15PPI=3P=II========================================5-*= Injection Date : 10/7/99 8:48:10 AM Seq. Line : 63 Sample Name : MFA-012 V i a l : 53 Acq. Operator : MTM Inj : 1 Acq. Method : C:\HPCHEM\~\METHODS\FOSESIM.M Pd 7.t Last changed : 10/6/99 1:56:00 P M by MTM Analysis Method : C:\HPCHEM\l\METHODS\1227-7OI.M SPA Last changed : 12/29/99 9:23:15 AM by MTM flM.vl.x (modified after loading) (Results are from a previously s SIM Analysis (ES-) for Et-FOSE-OH, MeFOSE-OH, THPFOS, and PFOS using 4 m m ~ 3 5 m mIlionex IonPac NG1 column, S/N 12879. MTM Page 69 of 71 BACK TO MAIN 3M Environmental Laboratory Report No.W1880 Batch Run # 12 of 50 Data File C:\HPCHEM\l\DATA\lO0699\MeFOSO62.D Signal 1: MSDl 427, EIC=426.7:427.7 RetTime Type - -[-m-i-nl- - 1 - - . - - - - 5.873 BE3 I T o ta1s w i t .hout Sample Name: MFA-011 Signal 2: MSDl 499, EIC=498.7:499.7 RetTime T y p e Iminl . Totals without ISTD(s) : 3.08284e-1 Signal 3: MSDl 630, EIC=629.7:630.7 RetTime Type Area Amt/Area Amount G r p Name - -[-m-i-n-]- 1 - - - - - - I - - - - - - - - - - l - - - -r-a-t-i-o- - l - - - - -C-P-P-b-l- l - - l - - - - - - - - - - - - - - - - - - 8.538 BB I 7.38896e5 1.00000 310.80000 EtFOSE-OH Totals without ISTD(s) : 0.00000 Signal 4: MSDl 616, EIC=615.7:616.7 Totals without ISTD(8) : 471.34921 Page 70 of 71 BACK TO MAIN 3M EnvironmentalLaboratory Report No.W1880 Batch Run # 13 of 50 Data File C:\HPCHEM\l\DATA\lO0699\MeFOSO63.D Signal 1: MSDl 427, EIC=426.7:427.7 RetTime Type Area Amt/Area Amount - -[-rn-i-n-l- I -.*---- ----------I------.---l---------- ratio ' [ppbl 5.872 PB I 4.24804e5 1,00000 402.60000 Totals without ISTD(9) : 0.00000 Sample Name: MFA-012 Signal 2 : MSDl 499, EIC=498.7:499.7 RetTime Type [minl Area Amt/Area Amount Totals without ISTD(s) : 1.07655e-2 Signal 3: MSDl 630, EIC=629.7:630.7 RetTime Type Area Amt/Area Amount Grp Name - -[r-n-in-l- - I - - . - - - - I - - - - - - - - - - l -r-a-t-i-o- L - - 1 -[P-P-b-l- - - - - - - - l - - J - - - - - - - - - - - - - - - - - - 8.535 PB I 7.26047e5 1.00000 310.80000 EtPOSE-OH Totals without ISTD(8) : 0.ooooo Signal 4: MSDl 616, EIC=615.7:616.7 RetTime Area Amt/Area Amount Grp Name - -[-mi-n-l- - I - - - - - - I - - - - - - - - - - l - -r-a-t-i-o- - - - l - - -[P-P-b-l- - - -I - - I - - - - - - - - - - - - - - - - - - 8.307 BV 1.49310e6 1.08429 693.02691 MeFOSE-OH Totals without ISTD(s) : 693.02691 Page 71 of 71